Background:Small Cajal body-specific RNAs (scaRNAs) are a specific subset of small nucleolar RNAs (snoRNAs) that have recently emerged as pivotal contributors in diverse physiological and pathological processes. However, their defined roles in carcinogenesis remain largely elusive. This study aims to explore the potential function and mechanism of SCARNA12 in bladder cancer (BLCA) and to provide a theoretical basis for further investigations into the biological functionalities of scaRNAs.Materials and Methods:TCGA, GEO and GTEx data sets were used to analyze the expression of SCARNA12 and its clinicopathological significance in BLCA. Quantitative real-time PCR (qPCR) and in situ hybridization were applied to validate the expression of SCARNA12 in both BLCA cell lines and tissues. RNA sequencing (RNA-seq) combined with bioinformatics analyses were conducted to reveal the changes in gene expression patterns and functional pathways in BLCA patients with different expressions of SCARNA12 and T24 cell lines upon SCARNA12 knockdown. Single-cell mass cytometry (CyTOF) was then used to evaluate the tumor-related cell cluster affected by SCARNA12. Moreover, SCARNA12 was stably knocked down in T24 and UMUC3 cell lines by lentivirus-mediated CRISPR/Cas9 approach. The biological effects of SCARNA12 on the proliferation, clonogenic, migration, invasion, cell apoptosis, cell cycle, and tumor growth were assessed by in vitro MTT, colony formation, wound healing, transwell, flow cytometry assays, and in vivo nude mice xenograft models, respectively. Finally, a chromatin isolation by RNA purification (ChIRP) experiment was further conducted to delineate the potential mechanisms of SCARNA12 in BLCA.Results:The expression of SCARNA12 was significantly up-regulated in both BLCA tissues and cell lines. RNA-seq data elucidated that SCARAN12 may play a potential role in cell adhesion and extracellular matrix (ECM) related signaling pathways. CyTOF results further showed that an ECM-related cell cluster with vimentin+, CD13+, CD44+, and CD47+was enriched in BLCA patients with high SCARNA12 expression. Additionally, SCARNA12 knockdown significantly inhibited the proliferation, colony formation, migration, and invasion abilities in T24 and UMUC3 cell lines. SCARNA12 knockdown prompted cell arrest in the G0/G1 and G2/M phase and promoted apoptosis in T24 and UMUC3 cell lines. Furthermore, SCARNA12 knockdown could suppress the in vivo tumor growth in nude mice. A ChIRP experiment further suggested that SCARNA12 may combine transcription factors H2AFZ to modulate the transcription program and then affect BLCA progression.Conclusions:Our study is the first to propose aberrant alteration of SCARNA12 and elucidate its potential oncogenic roles in BLCA via the modulation of ECM signaling. The interaction of SCARNA12 with the transcriptional factor H2AFZ emerges as a key contributor to the carcinogenesis and progression of BLCA. These findings suggest SCARNA12 may serve as a diagnostic biomarker and potential therapeutic target for the treatment of BLCA.
背景：小卡哈尔体特异性RNA（scaRNAs）是小核仁RNA（snoRNAs）的一个特定亚类，近年来在多种生理和病理过程中被证实具有关键作用。然而，其在肿瘤发生中的明确功能仍不清楚。本研究旨在探讨SCARNA12在膀胱癌（BLCA）中的潜在功能与机制，为进一步探索scaRNAs的生物学功能提供理论基础。 材料与方法：利用TCGA、GEO和GTEx数据库分析SCARNA12在BLCA中的表达及其临床病理学意义。采用实时定量PCR（qPCR）和原位杂交技术验证SCARNA12在BLCA细胞系和组织中的表达。通过RNA测序（RNA-seq）结合生物信息学分析，揭示SCARNA12表达差异的BLCA患者及SCARNA12敲低的T24细胞系中基因表达模式和功能通路的变化。运用单细胞质谱流式技术（CyTOF）评估受SCARNA12影响的肿瘤相关细胞群。此外，通过慢病毒介导的CRISPR/Cas9技术在T24和UMUC3细胞系中稳定敲低SCARNA12。分别通过体外MTT、克隆形成、划痕愈合、Transwell、流式细胞术实验以及体内裸鼠移植瘤模型，评估SCARNA12对细胞增殖、克隆形成、迁移、侵袭、凋亡、细胞周期及肿瘤生长的生物学影响。最后，通过RNA纯化染色质分离实验（ChIRP）进一步阐明SCARNA12在BLCA中的潜在作用机制。 结果：SCARNA12在BLCA组织和细胞系中均显著上调。RNA-seq数据表明，SCARNA12可能在细胞黏附和细胞外基质（ECM）相关信号通路中发挥潜在作用。CyTOF结果进一步显示，在SCARNA12高表达的BLCA患者中，富含波形蛋白（vimentin+）、CD13+、CD44+和CD47+的ECM相关细胞群显著富集。此外，敲低SCARNA12显著抑制T24和UMUC3细胞系的增殖、克隆形成、迁移和侵袭能力，并促使细胞阻滞于G0/G1期和G2/M期，同时促进细胞凋亡。体内实验表明，敲低SCARNA12可抑制裸鼠肿瘤生长。ChIRP实验进一步提示，SCARNA12可能通过结合转录因子H2AFZ调控转录程序，进而影响BLCA进展。 结论：本研究首次提出SCARNA12在BLCA中存在异常表达，并通过调控ECM信号通路阐明了其潜在的致癌作用。SCARNA12与转录因子H2AFZ的相互作用是促进BLCA发生发展的关键因素。这些发现表明，SCARNA12可能作为BLCA的诊断生物标志物和潜在治疗靶点。
肿瘤(癌症)患者之家